Solution to PS 8 #12

Only cells that have taken up the intact plasmids form colonies, because untransformed cells are sensitive to tertracycline.  There are two classes of plasmids: intact pBR322 formed by ligation of the ends cut by PstI, and a recombinant molecule formed by insertion of the PstI restriction fragment into the vector. Cells containing the intact vector are resistant to ampicillin, whereas thse containing the recombinant are sensitive to ampicillin, since the PstI site is within the amp8 gene.  These two types of cells can be ditinguished by transferring part of the colonies to another petri dish with medium containing ampicillin.  Cells from the colonies that fail to grow in the presence of ampicillin conatin the recombinant plasmid.



Return to Problem Set #8